Saliva sample collection with using Oragene®DNA kit

DNA extraction & purification

Gel electrophoresis

Experimental Outline

DNA quality and quantity assessment

Sequenom® MassARRAYGenotyping

3730xL DNA Analyzer Sanger Sequencing

1NanoDrop OD

1.0% Agarose Gel Electrophoresis

10,000bp

4,000bp

2,000bp

800bp

400bp

W1 W3D1 D2

50ng DNA /lane

W4D3 D4

Sample ID ng/µl A260 A280 A320

Estimated yield from 500ul sample (µg) (A260-A320) / (A280-A320)

D1 405.75 8.115 4.469 0.25 40.575 1.86D2 416.01 8.32 4.617 0.279 41.601 1.85W1 11.53 0.231 0.153 0.033 1.153 1.65D3 432.2 8.644 4.854 0.298 43.22 1.83D4 434.17 8.683 4.857 0.293 43.417 1.84W3 12.42 0.248 0.171 0.013 1.242 1.49W4 15.33 0.307 0.209 0.015 1.533 1.51

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Quality & Yield

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Corrected A260/A280

Estimated yieldFrom 500uLSaliva (µg)

Downstream Applications

Sequenom® MassARRAYSNP Genotyping

3730xl DNA Analyzer Sanger Sequencing

Sanger DNA Sequencing Results

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Sequenom® MassARRAY Results

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Percentage call rate(out of total 29 SNPs genotyped)

Sample W Sample D

Our Experience

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13 Extractions done7 IndividualsEasy to useHands-on time = 1.5-2.5 hrs

Good qualityWith individual variations

Data successfully generated for:

•Sanger DNA sequencing •Sequenom MassArray Genotyping

Acknowledgements

Lauren Scully

All GRC colleagues

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